The demand for petroleum based fuel has grown in an extensive way because of the increasing industrialization and the growth in transportation sector. This growth has a direct effect on the economy and remains a monopoly in the fuel trade. Bio-fuels are available in a larger volume across the world due to the advantage that they can be directly blended with the petroleum fuels and the results obtained are better when compared to the standard values of neat diesel fuel. Such kind of biofuels can be extracted from lemon fruit and its availability is abundance across globe. This work aims in introducing a new biofuel called as Lemon Essential Oil (LEO) which can be obtained through steam distillation process of lemon rinds. A 20% LEO is blended with diesel (LEO20+ Diesel80) and it is fed as fuel to the engine. The various properties of LEO are examined and they are compared to mineral diesel. The experimental investigation revealed that brake thermal efficiency of LEO20 is marginally higher than diesel at higher loads. Also significant reduction is observed for carbon monoxide, unburned hydrocarbon and smoke emissions. However, the oxides of nitrogen in LEO20 is comparatively higher than petroleum diesel at rated power output. Furthermore, the in-cylinder gas pressure of LEO20 is followed the similar trend with diesel fuel. In addition, higher heat release rate is also observed for LEO20 from the tested results. Hence, the present research work is showed that the 20% of Lemon Essential Oil could be partial substitute for conventional diesel engine in near future.